1,2,4-Benzenetricarboxylic acid, mixed decyl and octyl triesters

Endpoint summary

Additional information

1,2,4-Benzenetricarboxylic acid, mixed decyl and octyl triesters was tested for mutation in three histidine requiring strains (TA97, TA98 and TA100) of Salmonella typhimurium both in the absence and presence of metabolic activation by an Aroclor 1254 induced rat liver post-mitochondrial fraction (S9) according to OECD guideline 471. The substance failed to induce a two-fold increase in revertant numbers with any tester strain either in the absence or presence of S9, so was considered non-mutagenic in this assy.

The substance was assayed for the ability to cause chromosomal damage in cultured human lymphocytes, following in vitro treatment in the absence and presence of S9 metabolic activation according to Test method B.10 described in Council Regulation (EC) No. 440/2008 and OECD Guideline No. 473 (adopted July 1997).

Two independent experiments for chromosomal damage were performed. Following treatment with the test item, no statistically significant increase in the incidence of cells bearing aberrations, including or excluding gaps, was observed at any dose level of any sampling time. Statistically significant increases in the incidence of cells bearing aberrations (both including and excluding gaps) were seen following positive control treatments with Mitomycin-C and Cyclophosphamide, indicating the correct functioning of the assay system. It was concluded that 1,2,4 -Benzenetricarboxylic acid, decyl octyl ester did not induce chromosomal aberrations in human lymphocytes after in vitro treatment, under the reported experimental conditions. Also in a second chromosome aberration test the test item did not induce a statistically significant increase in the frequency of cells with chromosome aberrations, in either the absence or presence of a liver enzyme metabolizing system, in either of two separate experiments. Therefore also in the second test according to OECD 473 the test item can be considered to be non-clastogenic to human lymphocytes in vitro.

The test item 1,2,4 -Benzenetricarboxylic acid, decyl octyl ester was examined for mutagenic activity by assaying for the induction of 5 -trifluorothymidine resistant mutants in mouse lymphoma L5178Y cells after in vitro treatment, in the absence and presence of S9 metabolic activation, using a fluctuation method.

The study was designed to comply with the experimental methods indicated in: Test method B.17 "in vitro mammalian cell gene mutation test" described in Council Regulation (EC) No. 440/2008 and OECD Guideline for the testing of chemicals No. 476 (adopted July 1997). Both in the absence and presence of S9 metabolic activation, the test item was assayed at a maximum dose level of 2500 µg/mL and at a wide range of lower dose levels: 1250, 625, 313, 78.1, 39.1, 19.5 and 9.77 µg/mL. No relevant toxicity was observed at any dose level at any sampling time, in the absence and presence of S9 metabolism.

Based on the toxicity results obtained in the preliminary assay, two independent assays for mutation to 5 -trifluorothymidine resistance were performed using the following dose levels and treatment times:

Assay No. 1: 156, 313, 626, 1250 and 2500 µg/mL, 3 hours treatment with and without metabolic activation

Assay No. 2: 156, 313, 625, 1250 and 2500 µg/mL, 24 hours treatment without meatbolic activation; resp. 875, 1138, 1479, 1923 and 2500 µg/mL, 3 hours treatment with metabolic activation. No relevant increases in mutant frequencies were observed following treatment with the test item, in the absence or presence of S9 metabolism. Marked increases were obtained with the positive control treatments indicating the correct functioning of the assay system.

It was concluded that 1,2,4 -Benzenetricarboxylic acid, decyl octyl ester does not induce mutation at the TK locus of L5178Y mouse lymphoma cells in vitro in the absence or presence of S9 metabolic activation, under the reported experimental conditions.

All available data clearly demonstrate the lack of genotoxic effects.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Data from various tests clearly demonstrate that the material is not genotoxic and therefore the substance needs not to be classified.